Search Results (370)

Repetitive temperature fluctuations during transportation and storage promote ice crystal formation in salmon flesh, leading to protein denaturation, lipid oxidation, and quality loss. Tuna skin, a major by-product of tuna processing, is a potential source of antifreeze peptides (AFPs) but remains underutilized. This study examined the cryoprotective effects of tuna skin-derived AFPs on salmon cubes subjected to repeated freeze–thaw cycles. Cubes treated with AFPs from three groups of protein hydrolysates prepared using trypsin, pepsin, or neutral protease were evaluated for texture, color, water holding capacity (WHC), volatile odor profiles, protein conformation, biochemical indices, and microstructure. AFP treatment improved textural properties, maintained color stability, and reduced thawing, cooking, and centrifugal losses. The neutral protease-treated group exhibited the optimal cryoprotective ability and it also limited aldehyde and sulfide accumulation, preserved the retention rate of α-helix structure at 49% which was higher than 39% in controls, and enhanced Ca²⁺-ATPase activity to 1.75 μmol Pi·mg⁻¹·h⁻¹ with a 45.8% increase compared to controls, and significantly inhibited protein and lipid oxidation. Microstructural analysis showed compact fibers and intact sarcolemma in the neutral protease-treated group samples, contrasting with severe disruption in controls. This study showed that tuna skin AFPs mitigate freeze–thaw damage in salmon cubes by stabilizing proteins and reducing oxidative deterioration, highlighting their potential as natural, healthy cryoprotectants for seafood preservation, meeting the growing demand of the food industry for clean-label, low-calorie preservation solutions, while advancing the circular economy of aquatic processing via the valorization of tuna skin by-products for high-value seafood applications. Full article

Collagen is the primary protein in the extracellular matrix of human cells and the body and is essential for cell structure and function. Here, for the first time, we report a method for producing recombinant triple-helical collagen type III (rhCOL3) in transgenic tobacco as a bioreactor. We constructed a pMDV-COL3A1 vector containing the human type III collagen gene COL3A1, as well as a pMDV-COL3A1:5E vector that coexpressed COL3A1 and the enzymes required for its posttranslational modification. These two vectors were used to transform tobacco genetically. The COL3A1 gene was successfully coexpressed in tobacco plants with four enzymes that promote its posttranslational modification. The transcriptional level of COL3A1 in the transgenic lines coexpressing posttranslational modification genes was greater than that in the transgenic lines expressing only COL3A1. The enzyme-modified recombinant collagen was subsequently purified from a COL3A1:5E transgenic line. Our experimental results demonstrated that the terminal propeptides of plant-derived rhCOL3 can be correctly cleaved through the enzymatic hydrolysis of procollagen by coexpressed procollagen C proteinase (PCP) and procollagen N proteinase (PNP). The plant-derived rhCOL3 was thermally stable because the purified peptide chains can form a triple helix structure. Experiments have shown that plant-derived rhCOL3 has biological activity. In this study, functional recombinant full-length mature type III collagen with a triple-helix structure was successfully expressed in tobacco, providing a foundational plant-made material for future applications of collagen in human skin and bone repair in regenerative medicine. Full article

Growing demand for low-sodium surimi products has driven the search for safe salt alternatives. Anionic peptides (APPs) and cationic peptides (CPPs) were isolated from sheepskin collagen via Diethylaminoethyl (DEAE) chromatography. CPPs contained higher arginine (46.11%) and lysine (4.64%) than APPs (40.57% and 3.99%, respectively), while APPs were enriched in acidic amino acids like glutamic acid (3.88%). Comprehensive evaluations of low-salt silver carp surimi gels showed both peptides significantly improved gel strength and water-holding capacity (WHC). The water-holding capacity increased from 60.68% in the blank control group to 74.31% in the CPP-treated group, while that in the APP-treated group was 66.86%. Cooking loss was significantly reduced, decreasing from 40.64% in the blank control group to 28.57% in the CPP-treated group and 34.52% in the APP-treated group. The samples achieved a quality comparable to that of the NaCl-supplemented group, with CPP outperforming APP in terms of hardness and gel network density. The LF-NMR confirmed enhanced water retention by reducing free water (T₂₂) and increasing bound water (T₂b). The FTIR indicated a conformational shift from α-helix to β-sheet, and the SEM revealed denser networks with fewer large voids. The SDS-PAGE demonstrated enhanced myosin heavy chain (MHC) cross-linking, more pronounced in the CPP-treated samples. CPPs exerted stronger electrostatic attraction with negatively charged surimi proteins (isoelectric point 5.5), while APPs chelated Ca²⁺ to activate transglutaminase. These findings validate APPs and CPPs as promising salt substitutes, enabling low-sodium surimi production and high-value utilization of sheepskin by-products. Full article

Haliotis discus hannai meat has a firm texture that makes it difficult to chew and swallow, so tenderizing is necessary. Ultrasonic treatment and papain enzyme processing are used to reduce the hardness of abalone meat. This study tests physicochemical indicators and protein changes to assess meat quality and protein alterations. The maximum reduction in hardness of raw meat reached 60.58%, while heat-treated abalone meat achieved 61.13%, with free water and bound water converting to immobile water. The L* value of the meat decreased, while the a* and b* values increased. In raw meat, the content of TCA-soluble peptides increased with increasing treatment intensity. However, in heat-treated meat, this peptide content decreased with rising temperature. Muscle fiber filament breaks and pore numbers increased. The BPB binding content showed a negative correlation with the percentage of α-helix. Total sulfhydryl and free amino groups in raw meat decrease with increasing treatment intensity; both parameters in cooked meat decrease with rising temperature. Changes in tertiary protein structure cause alterations in fluorescence intensity, with secondary structure shifting from α-helix to β-sheet conformation. The results suggested that ultrasonic and papain treatments can induce structural alterations in proteins. This leads to protein cleavage and depolymerization, collectively resulting in softening of abalone meat texture and redistribution of internal moisture. These processes result in softening of abalone meat and redistribution of internal moisture. This study provides a theoretical basis for developing abalone tenderizers. Full article

Therapeutic proteins face a critical pharmacokinetic challenge: rapid clearance from circulation limits their clinical efficacy. Albumin-binding domains (ABDs) offer an elegant solution by enabling therapeutic proteins to “hitchhike” on serum albumin’s favorable 19-day half-life through FcRn-mediated recycling. Clinical validation through approved therapeutics like ozoralizumab demonstrates the success of this approach, with preclinical studies showing fusion to an ABD extended half-life to 18 days. This review provides an analysis of ABD-fusion protein design, integrating structural biology, computational prediction, and rational engineering principles. We catalog the major classes of albumin-binding modalities, including bacterial three-helix bundle domains, engineered peptides, antibody-derived binders, and alternative scaffolds, comparing their binding properties, size contributions, cross-species reactivity, and production cost. Critical examination of linker architectures reveals that flexible glycine-serine linkers (particularly the widely successful (GGGGS)₃ motif) provide optimal balance between domain independence and molecular economy, though linker choice profoundly influences not only spatial separation but also binding affinity, folding, stability, and pharmacokinetics. We evaluate the utility and limitations of the structure prediction tools for ABD-fusion design. We establish practical guidelines for integrating computational screening with experimental validation. This review provides protein engineers and synthetic biologists with a comprehensive framework for rational design of albumin-binding therapeutics, emphasizing the synergistic integration of structural insight, computational prediction, and systematic experimental validation to accelerate development of next-generation long-acting biotherapeutics. Full article

The broadly neutralizing monoclonal antibody 3D1 potently neutralizes SADS-CoV by targeting a conserved epitope within the heptad repeat 1 (HR1) domain of the viral spike protein. Structural and biophysical analyses demonstrate that 3D1 binds with high affinity to a specific linear β-turn motif (residues A804–N809) in HR1. High-resolution crystallography reveals that this motif sits within a deep, electrostatically complementary paratope groove. Critically, 3D1 binding competitively inhibits the essential interaction between HR1 and HR2. Notably, its recognition is not dependent on HR1’s native helical conformation, as it maintains strong binding to conformationally constrained, stapled helical peptides. Collectively, the data indicate that 3D1 neutralizes by capturing a pre-hairpin intermediate state of HR1—a transition state between prefusion and postfusion forms—thereby sterically blocking the formation of the stable postfusion six-helix bundle that is essential for membrane fusion. This work defines a precise, structure-dependent neutralizing epitope and elucidates a mechanism of action that involves trapping a key fusion intermediate, offering a valuable template for the design of broad-spectrum coronavirus therapeutics. Full article

Rice bran, a nutrient-rich by-product of rice milling, is an underutilized resource in sustainable crop utilization. This study aimed to investigate the characteristics, total phenolic content, and antioxidant activities of rice bran protein hydrolysates (RBPHs) produced using proteases from Bacillus licheniformis (RBPH-B) and α-chymotrypsin (RBPH-C), along with their protein fractions (F1; >100 kDa, F2; 10–100 kDa, F3; 1–10 kDa, F4; <1 kDa). Molecular weight, color, surface hydrophobicity, secondary structure, total phenolic content, and antioxidant activities of the hydrolysates were assessed. Both enzymatic hydrolysis and ultrafiltration reduced molecular weight and surface hydrophobicity, enhanced lightness, and increased α-helix content. Among all samples, the <1 kDa peptide fraction derived from α-chymotrypsin hydrolysis (RBPH-C-F4) exhibited the strongest antioxidant activity, with the lowest EC₅₀ values for ABTS (0.94 mg/mL) and DPPH (210 µg/mL), as well as the highest inhibition of metal chelating activity (1.35 mmol EDTA/g sample) and linoleic peroxidation (90.62%). Enzymatic hydrolysis enhanced total phenolic content compared with native rice bran protein. These findings highlight the potential of rice bran-derived peptides as antioxidant candidates and indicate that further validation in food systems is required. Full article

The increasing crisis of antimicrobial resistance requires innovative therapeutic strategies that can overcome the limitations of conventional antibiotics. Based on our previous finding that AP10 (a derivative of AP29) possesses antimicrobial activity but lacks thermal stability, we rationally redesigned ten new AP10 analogues to enhance functional robustness while maintaining efficacy. Among these, AP10RW is identified as the optimal candidate due to its exceptional broad-spectrum activity against both drug-sensitive and multidrug-resistant (MDR) bacterial pathogens. Structural analysis reveals that AP10RW adopts an environmentally responsive conformation, transitioning from random coil to amphiphilic α-helix in membrane-mimicking environments, while demonstrating remarkable stability under challenges including serum exposure, varying pH, high salt concentrations, and thermal stress. Mechanistic studies indicate that AP10RW exerts its effects through multiple bactericidal mechanisms involving initial high-affinity binding to bacterial characteristic molecules (LTA, LPS and PGN), followed by rapid membrane depolarization, ultrastructural damage and the induction of lethal oxidative stress. Notably, this potent antimicrobial efficacy is coupled with exceptional biosafety, demonstrating little hemolysis and negligible cytotoxicity against mammalian cells. This systematic optimization represents a significant advancement in antimicrobial peptide engineering. We have successfully transformed a thermally unstable peptide into a robust therapeutic candidate and positioned AP10RW as a promising clinical candidate for addressing the growing threat of multidrug-resistant infections. Full article

An upgraded GAFF2 force field has been used to simulate two fluorinated alcohols, TFE and HFIP, in aqueous solutions at several concentrations. The same force field has also been employed to simulate a 26-residue amphiphilic peptide in several cosolvent/water mixtures to verify and clarify its efficacy in stabilizing the secondary structure. The calculated thermodynamic and structural properties are in agreement with experimental findings. The force field allows a correct description of the secondary structure and affords an accurate characterization of the spatial organization of cosolvent molecules around the peptide. Full article

To identify pancoronaviral inhibitors, we sought to identify peptides that bound the evolutionarily conserved SARS-CoV-2 spike fusion peptide (FP). We screened the NEB PhD-7-mer random combinatorial phage display library against FP, synthesised as a D-peptide, to identify peptides from the L-library to be synthesised as proteolytically resistant D peptides. We selected the top ten peptides that were not seen in another published screen with this library, as these were more likely to be specific. All ten D-peptides had no impact on the infection of Vero-E6/TMPRSS2 cells by SARS-CoV-2. Screening of a proteomic-derived phage display library from the disordered regions of human proteins identified two overlapping 14mer peptides from a region of OTUD1. While a synthetic peptide based on their sequences failed to markedly inhibit viral entry, molecular dynamics structural modelling highlighted a stable binding mode where positive residues on one side of the OTUD1 helix interacted with hydrophobic residues of the FP triple-helical wedge. Thus, while the two phage display strategies failed to yield peptide sequences that are themselves strong inhibitors of viral infection, they led to the development of a computational model that can underpin future designs of potential pancoronaviral FP disruptors. Full article

Viral fusion proteins are indispensable mediators of viral entry that orchestrate the fusion of viral and host membranes, making them primary targets for antiviral interventions. Class I fusion proteins, displayed on the surface of enveloped viruses (such as HIV-1, RSV, SARS-CoV-2, Nipah, influenza, and Ebola viruses), share conserved structural features, including the fusion peptide or loop and heptad repeat regions. These elements are essential for the formation of the post-fusion six-helix bundle during membrane fusion. Peptide inhibitors that mimic heptad repeat motifs have consequently emerged as an effective strategy for blocking the fusion process. This review summarizes design strategies for such inhibitors and highlights how sequence and structural insights have enabled their optimization via α-helical stabilization, hydrocarbon stapling, lactam bridges, lipid conjugation, macrocyclization, and multivalency. Using representative examples across major viral systems, this review illustrates how these strategies have led to the development of potent, stable, and even broad-spectrum antiviral peptides. This review provides insights to guide the rational design of next-generation peptide-based fusion inhibitors targeting viral membrane fusion. Full article

Collagen, the most abundant structural protein in animals, is fundamental for tissue integrity and regeneration. Conventional mammalian sources face limitations related to sustainability, safety, and ethical concerns, underscoring the need for alternative biomaterials. Marine organisms, particularly jellyfish, offer a promising eco-friendly collagen source. In this study, collagen and collagen-derived peptides were extracted from the cnidarian Physalia physalis and biochemically characterized. Circular dichroism demonstrated partial loss of triple-helix structure, while SDS-PAGE revealed type I collagen related α-chains together with low-molecular-weight fragments. The hydrolyzed collagen fractions exhibited keratinocyte and fibroblast cytocompatibility and increased keratinocyte migration. Moreover, P. physalis-derived peptides modulated inflammatory cytokine release in lipopolysaccharide-stimulated macrophages reducing tumor necrosis factor (TNF)-α by 38% and increasing interleukin (IL)-10 by 29%. Based on these results, a stable bioactive serum formulation incorporating P. physalis collagen peptides was developed. Overall, this work demonstrates that bioactive peptides from P. physalis possess immunomodulatory and regenerative potential and represent a promising new marine resource for cosmetic applications. Full article

Marine algae are a prolific bioactive peptide source with a broad pharmacological potential. We characterized MP28, a cationic peptide isolated from the green alga Bryopsis plumosa. Structural modeling indicated a predominantly amphipathic α-helix (residues 3–16) flanked by flexible termini and stabilized by intramolecular disulfide bonds, a motif typical of membrane-active anticancer peptides. Functionally, MP28 demonstrated potent activity against non-small-cell lung cancer cell lines (A549, H460, H1299) without affecting non-tumorigenic lung fibroblasts (MRC-5). In vitro, MP28 decreased cell viability and clonogenic growth and suppressed migration and invasion in a dose-dependent manner. Flow cytometry revealed increased early/late apoptotic fractions, accompanied by caspase-9 activation, consistent with engagement of the intrinsic apoptotic pathway. In a mouse xenograft model, MP28 treatment significantly reduced tumor size compared with that of controls. Collectively, MP28 may be a potent anticancer peptide that exhibits selective cytotoxicity and low toxicity toward normal cells. Full article